OpenWetWare:Getting started: Difference between revisions

Revision as of 14:34, 10 August 2015

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   <a class="navbar-brand" href="http://openwetware.org/wiki/Biomod/2014/OhioMOD">OhioMOD</a>
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     <li ><a href="http://openwetware.org/wiki/Biomod/2014/OhioMOD">Home</a></li>
     <li><a href="http://openwetware.org/wiki/Biomod/2014/OhioMOD/project">Background</a></li>

<li><a href="http://openwetware.org/wiki/Biomod/2014/OhioMOD/results">Project and Results</a></li>

       <a href="#" class="dropdown-toggle" data-toggle="dropdown">Materials and Methods <b class="caret"></b></a>
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         <li><a href="http://openwetware.org/wiki/Biomod/2014/OhioMOD/protocols">Protocols</a></li>
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         <li><a href="http://openwetware.org/wiki/Biomod/2014/OhioMOD/LabNotebook">Experiment Schedule</a></li>
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<li><a href="http://openwetware.org/wiki/Biomod/2014/OhioMOD/team">Team</a></li> <li><a href="http://openwetware.org/wiki/Biomod/2014/OhioMOD/sponsors">Acknowledgement</a></li> <li><a href="http://www.giveto.osu.edu/igive/OnlineGiving/fund_results.aspx?fund=314139" target="_blank">Give To OhioMOD</a></li>

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</style>  </head> <body> <div id="logo"> <img src="http://openwetware.org/images/e/ed/OHIOMODLOGO2.png" alt="OhioMOD"> <iframe src="//www.facebook.com/plugins/like.php?href=https%3A%2F%2Fwww.facebook.com%2FOhioMod&width&layout=standard&action=like&show_faces=true&share=true&height=80" scrolling="no" frameborder="0" style="border:none; overflow:hidden; height:80px;" allowTransparency="true"></iframe> </div> <div id="abstract"> <h1>Welcome to OhioMOD</h1> <p1>OhioMOD is Ohio State's undergraduate biomolecular design team. The team participates in the BIOMOD international competition each year at Harvard University. The 2014 project will use a folded DNA structure to control miRNA gene regulation in cancer.</p1> <h1>Abstract</h1> <p2>MicroRNAs (miRs) are short, non-coding RNAs that regulate gene expression and control essential cellular processes, such as cell cycle progression and apoptosis, through interactions with messenger RNA. Dysregulation of miR expression levels can promote tumorigenesis either by elevating oncogene or silencing tumor suppressor gene levels implicating miRs as an attractive therapeutic target. One therapeutic approach is to decrease levels of miRs that target tumor suppressor genes such as PTEN to ultimately promote apoptosis by complementary base pair antagonism via antisense DNA, but limitations exist in regard to antisense DNA cellular delivery. A previous study demonstrated that DNA nanostructures functionalized with small interfering RNA (siRNA) underwent efficient cellular entry and executed targeted gene suppression suggesting strong potential for DNA nanostructures as a nucleotide delivery vehicle. Here, we report the fabrication of a rod-like and a block-like DNA nanostructure incorporated with DNA overhangs capable of binding and sequestering miR-21, a PTEN targeting miR that is overexpressed in a variety of malignancies including chronic lymphocytic leukemia (CLL). Proper assembly was confirmed via gel electrophoresis and transmission electron microscopy. Incorporation and functionality of DNA overhangs complementary for miR-21 was confirmed via fluorescent gel imaging. In addition, DNA nanostructures were shown to enter the endolysosomal pathway as revealed via fluorescent microscopy. Furthermore, miR-21 complementary DNA overhang functionalized structures induced marked decreases in OSU-CLL cell viability relative to scrambled control overhang nanostructures as shown via fluorescent microscopy. Future work will directly investigate expression levels of PTEN mRNA and protein using quantitative real time PCR and immunoblot analysis respectively. Our current findings suggest that the complementary miR-21 DNA overhang functionalized DNA nanostructures effectively sequester miR-21 to induce cytotoxicity in OSU-CLL cells. Our results represent a promising antisense DNA delivery therapeutic approach in a CLL leukemia model with implications to a variety of human malignancies.</p2> <h1>Project Video</h1> <br> </div> <div id="video"> <center> <iframe width="660" height="415" src="//www.youtube.com/embed/OzRtxW4YK2c" frameborder="0" allowfullscreen></iframe> </center>

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<td id="email">Contact us at <a href="mailto:theohiomod@gmail.com"><font color="white">theohiomod@gmail.com</a></td>

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OpenWetWare:Getting started: Difference between revisions

Revision as of 14:34, 10 August 2015

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-{{OpenWetWare:Getting started/Introduction}} <!-- PLEASE DO NOT MAKE EDITS TO THIS PAGE! If you would like to make test edits, please refer here: http://openwetware.org/wiki/Sandbox -->
+{{OhioMOD2014}}
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-== Introduction ==
-A hematopoietic stem cell is a stem cell that is derived from the bone marrow marrow or the blood of a subject. These stem cells are pluripotent and thus have the ability to be transformed into any other type of blood cell or immune cell. Their role within the blood is to keep the body constantly replenished with blood as the blood cells must be replaced every day. There are two different types of hematopoietic stem cells, long term and short term. However it is extremely difficult for researchers to differentiate between the two different types of stem cells once they are removed from the blood or bone marrow. The difference between the two types of cells are that long term can regenerate indefinitely while short term stem cells cannot renew themselves over a long period of time.
-[[Image:Stem cell.jpg]]
+</style>
+<!--css stuff ends here-->
+</head>
+<body>
+<div id="logo">
+<img src="http://openwetware.org/images/e/ed/OHIOMODLOGO2.png" alt="OhioMOD">
+<iframe src="//www.facebook.com/plugins/like.php?href=https%3A%2F%2Fwww.facebook.com%2FOhioMod&amp;width&amp;layout=standard&amp;action=like&amp;show_faces=true&amp;share=true&amp;height=80" scrolling="no" frameborder="0" style="border:none; overflow:hidden; height:80px;" allowTransparency="true"></iframe>
+</div>
+<div id="abstract">
+<h1>Welcome to OhioMOD</h1>
+<p1>OhioMOD is Ohio State's undergraduate biomolecular design team.  The team participates in the BIOMOD international competition each year at Harvard University.  The 2014 project will use a folded DNA structure to control miRNA gene regulation in cancer.</p1>
+<h1>Abstract</h1>
+<p2>MicroRNAs (miRs) are short, non-coding RNAs that regulate gene expression and control essential cellular processes, such as cell cycle progression and apoptosis, through interactions with messenger RNA. Dysregulation of miR expression levels can promote tumorigenesis either by elevating oncogene or silencing tumor suppressor gene levels implicating miRs as an attractive therapeutic target. One therapeutic approach is to decrease levels of miRs that target tumor suppressor genes such as PTEN to ultimately promote apoptosis by complementary base pair antagonism via antisense DNA, but limitations exist in regard to antisense DNA cellular delivery. A previous study demonstrated that DNA nanostructures functionalized with small interfering RNA (siRNA) underwent efficient cellular entry and executed targeted gene suppression suggesting strong potential for DNA nanostructures as a nucleotide delivery vehicle. Here, we report the fabrication of a rod-like
+and a block-like DNA nanostructure incorporated with DNA overhangs capable of binding and sequestering miR-21, a PTEN targeting miR that is overexpressed in a variety of malignancies including chronic lymphocytic leukemia (CLL). Proper assembly was confirmed via gel electrophoresis and transmission electron microscopy. Incorporation and functionality of DNA overhangs complementary for miR-21 was confirmed via fluorescent gel imaging. In addition, DNA nanostructures were shown to enter the endolysosomal pathway as revealed via fluorescent microscopy. Furthermore, miR-21 complementary DNA overhang functionalized structures induced marked decreases in OSU-CLL cell viability relative to scrambled control overhang nanostructures as shown via fluorescent microscopy. Future work will directly investigate expression levels of PTEN mRNA and protein using quantitative real time PCR and immunoblot analysis respectively. Our current findings suggest that the complementary miR-21 DNA overhang functionalized DNA nanostructures effectively sequester miR-21 to induce cytotoxicity in OSU-CLL cells. Our results represent a promising antisense DNA delivery therapeutic approach in a CLL leukemia model with implications to a variety of human malignancies.</p2>
+<h1>Project Video</h1>
+<br>
+</div>
+<div id="video">
+<center>
+<iframe width="660" height="415" src="//www.youtube.com/embed/OzRtxW4YK2c" frameborder="0" allowfullscreen></iframe>
+</center>
-== Motivation ==
+<A HREF="#top"><img src="http://openwetware.org/images/4/46/Arrow_up.png" id="fixedbutton"height="40" width="40"></A>
-Hematopoietic stem cells can be taken from a donor patient and inserted into a patient with a compromised immune system or bone marrow. This process is called hematopoietic stem cell transplantation, and the donor stem cells are inserted via an IV into the patient. This transplantation method can be used to treat patients who have diseases such as neuroblastoma or Non-Hodgkin Lymphoma. However there are numerous potential side effects including infection as well as Graft-versus-host disease.
+</br><a href="http://openwetware.org/index.php?title=Biomod/2014/OhioMOD&action=edit">Edit Main Page</a>
-Another use for hematopoietic stem cells are to use them to induce tolerance in solid organ grafts. By introducing these stem cells into the organ that’s been transplanted, the autoimmune response to that new organ is reduced or eliminated. This effect was examined in newborn mice, where transplanted organs were accepted by the host mice’s body for the remainder of its life and no autoimmune response to the new organ observed (Billingham et. Al). The reason for the acceptance of the donated organ is due to a lower negative T cell response, essentially meaning that T cells won’t recognize the new organ as an invader and attack it.
-[[Image:Boom.png]]
+<a href="http://info.flagcounter.com/4FJ3"><img src="http://s06.flagcounter.com/count/4FJ3/bg_FFFFFF/txt_000000/border_CCCCCC/columns_8/maxflags_12/viewers_3/labels_1/pageviews_1/flags_0/" alt="Flag Counter" border="0"></a>
+</div>
-== Future Work/Complications ==
+</body>
-Research has been done on whether or not hematopoietic stem cells can be used to heal other parts of the body rather than blood or bone marrow. There is evidence to suggest that hematopoietic stem cells tend to gravitate towards injured areas of the body and heal damaged tissues there. There is also the potential of these stem cells being turned other types of cells such as bone cells or brain cells. However the viability of this method is still in question as the research into this is still fairly young and as yet unexplored although the potential is still there.
+</html>
-One of the key obstacles to further work in this area is the limited supply of hematopoietic stem cells as well as the very large numbers needed by patients for each treatment. This is due to the host’s body tendency to attack the donated cells. Another obstacle of these stem cells is the inability of scientists to be able to differentiate between long term and short term hematopoietic stem cells, as only the long term stem cells are useful for growing cultures. These long term cells are also fairly rare, only occurring 1:10,000 normal red blood cells.
-== References ==
+{{OSUFooter}}
- ==Toshihisa Tsuruta (2013). Recent Advances in Hematopoietic Stem Cell Gene Therapy, Innovations in Stem Cell Transplantation, Prof. Taner Demirer (Ed.), ISBN: 978-953-51-0980-8, InTech, DOI: 10.5772/53587. Available from: http://www.intechopen.com/books/innovations-in-stem-cell-transplantation/recent-advances-in-hematopoietic-stem-cell-gene-therapy
+{{OpenWetWare:Getting started/Introduction}} <!-- PLEASE DO NOT MAKE EDITS TO THIS PAGE! If you would like to make test edits, please refer here: http://openwetware.org/wiki/Sandbox -->
- ==Regenerative Medicine. Department of Health and Human Services. Chapter 2: Bone Marrow (Hematopoietic) Stem Cells. August 2006. http://stemcells.nih.gov/info/scireport/2006report.htm
- ==Hematopoietic Stem Cells . In Stem Cell Information [World Wide Web site]. Bethesda, MD: National Institutes of Health, U.S. Department of Health and Human Services, 2011 [cited Wednesday, January 28, 2015] Available at <http://stemcells.nih.gov/info/scireport/pages/chapter5.aspx>